Rubus glaucus is, perhaps, one of the most desirable fruits in the market. their diversity of uses has become one of the promising crops of the region. Although the specie has been the focus of many investigations aimed at the spread, there is a gap in information that contributes to their cytogenetic characterization. This research was focused on the standardization of the technique of crushed roots or squash for the counting of chromosomes in somatic cells. For a pre-treatment, it was used 8-Hydroxyquinoline during 4 hours at room temperature. The solution was fixed with Farmer for 24 hours at 5 oC. There were two types of hydrolysis: an acidic and an enzymatic hydrolysis. The Staining was carried out with Acetocarmin. The number of chromosomes found in Rubus glaucus was 28 chromosomes (tetraploidy). There were not found differences between singular sources grown in the coffee belt.